University Of Minnesota

NIH Research Projects · FY 2024 · 2024-08

PROJECT SUMMARY CANDIDATE: Daniel H. Craighead, PhD, is a postdoctoral fellow training in integrative physiology at the University of Colorado (CU) Boulder. In this K01 application, Dr. Craighead aims to determine the efficacy of high-resistance inspiratory muscle strength training (IMST), a novel and time-efficient lifestyle intervention, for lowering systolic blood pressure (SBP) and improving endothelial, cerebrovascular, and cognitive function. His immediate goal is to acquire the research training and professional skills necessary to transition to an independent, extramurally funded investigator. His long-term goal is to establish his own research program with a focus on identifying novel, evidence-based lifestyle interventions that lower blood pressure and prevent or delay the development of blood pressure-associated comorbidities and chronic diseases. CAREER DEVELOPMENT PLAN: Dr. Craighead's career development plan consists of: 1) acquiring new skills to assess cerebrovascular and cognitive function to support his proposed research plan; 2) train in new cellular and molecular techniques to assess changes in oxidative stress and nitric oxide-bioavailability; and 3) professional skill development through coursework; attendance/presentations at weekly journal clubs, CU seminars, and national scientific meetings; and regular interactions with his mentor team. ENVIRONMENT: The environment for Dr. Craighead's training plan will be outstanding. Dr. Craighead's primary mentor, Dr. Douglas Seals, and co-mentor, Dr. Michel Chonchol, are internationally recognized, NIH-funded scientists with strong records of successful mentoring in translational biomedical research. Co-mentor, Dr. Fiona Bailey, performed the pioneering work on high-resistance IMST, making her the foremost expert of this novel lifestyle intervention. Consulting mentor Dr. Zhiying You is Senior Biostatistician in the Department of Medicine at the CU Anschutz Medical Campus and regularly provides mentoring/consulting to trainees and faculty conducting clinical trials. Consulting mentor Dr. Philip Ainslie is the Canada Research Chair in Cerebrovascular Physiology with extensive experience assessing cerebrovascular function. Consulting mentor Dr. Brianne Bettcher is an Assistant Professor at the CU Anschutz Medical Campus and expert in assessing cognitive function. RESEARCH: Above-normal SBP is a highly prevalent condition, afflicting the majority of adults over age 50. Above-normal SBP increases the risk for developing cardiovascular diseases, stroke, cognitive decline, and other chronic conditions. This increased risk is largely attributable to the development of vascular dysfunction in the peripheral and cerebral arteries, secondary to an increase in oxidative stress and consequent reduction in nitric oxide-bioavailability. Healthy lifestyle practices are first-line therapies to lower SBP and decrease disease risk; however, adherence to these practices is poor due to their time-intensive nature. The proposed research will test the efficacy of time-efficient, high-resistance IMST for promoting adherence, lowering SBP, and improving endothelial, cerebrovascular, and cognitive function in adults with above-normal SBP.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Alzheimer’s disease (AD) is the most common type of dementia, afflicting approximately 5.7 million people, or 60–80% of cases of dementia in the United States, and is predicted to triple to 16 million by the year 2050. Diagnosis is made after the age of 65 in 90-95% of patients, with the remaining 5-10% considered early-onset. Symptoms usually begin with difficulty learning new facts or recalling recent memories. In the middle stage of AD progression, language becomes more difficult, memory continues to deteriorate with long-term memory now also being affected, patients may not recognize close relatives, and will need assistance with most activities of daily living. During the late stage of AD, language is eventually lost, muscle atrophies to the degree that patients cannot get out of bed or feed themselves, and pneumonia or infection of pressure ulcers usually leads to death. Thus, AD is clearly a significant health problem. The biochemical basis for AD pathophysiology is thought to be the aggregation of extracellular amyloid β (Aβ) plaques and intracellular neurofibrillary tangles of hyperphosphorylated tau protein. These markers correlate with the severity of clinical signs and symptoms of the disease, and it is thought that they may be directly neurotoxic. However, a more recent paradigm is that these aggregates of misfolded proteins may lead to a state of chronic neuroinflammation, and that this inflammatory state is responsible for loss of neuronal function and neuron death. This inflammatory state is achieved through activation of the innate microglia, as well as the relatively understudied (in this context) activation of adaptive T lymphocytes. Regulatory T cells (Tregs) are a subset of T cells that dampen immune responses and promote tissue repair after injury in various tissues including the brain. Recently, regulatory T cells have been suggested to play an important role in AD. However, important gaps in our understanding of Tregs in AD remain: What types of Tregs are found in the brain in AD mice relative to wildtype mice? and, What role do Tregs play in the progression of AD? I will address this knowledge gap in two specific aims. In Aim 1, I will characterize the types of Tregs and their spatial localization in the aging WT and AD brains, while in Aim 2 I will establish the effect of increasing or decreasing Treg numbers or function. My hypothesis is that brain-specific Tregs play a critical role in preventing or delaying AD progression, and that increasing their number and function will lead to improved outcomes in AD. Successful completion of the aims of this grant will provide insight into the role of Tregs in AD. These studies will enhance my training in computational and hypothesis generating experiments (Aim 1), as well as hypothesis driven functional studies (Aim 2).

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ABSTRACT This K23 Career Development Award is designed to provide the training needed for the PI to achieve her long-term career goal of conducting independent, programmatic intervention research in developmental populations. The training will emphasize gaining expertise in higher-intensity, multi-method, within-subject data collection and analysis. This award builds on the PI’s emerging experience in tic disorders and pediatric behavioral interventions, and her ability to quickly learn and apply advanced statistical methods. The award will extend the PI’s training through the following short-term training goals: 1) multi-method data collection and integration (electronic momentary assessment [EMA], wearable devices, neurocognitive tasks), 2) leading and designing pediatric clinical trials, 3) managing and analyzing large, multilevel datasets, and 4) career development and contribution to the field. The PI has developed a training plan to accomplish these goals in concert with her mentors, a team of leading experts in the fields of psychiatry and psychology, who will closely monitor training through regular meetings. The highly structured training plan also includes a set of formal coursework and workshops for each training goal to complement the hands-on experience the PI will gain from leading the research project. The objective of this proposal is to comprehensively map symptom change across time and during a behavioral intervention for youth with Persistent Tic Disorders (PTDs). PTDs affect approximately 1% of the population, can cause significant disability, have high rates of comorbidity, and are associated with a four-fold increase in suicide risk. Research has established that tic symptoms and their change over time are highly idiographic. However, first-line, evidence-based, existing interventions are “one-size-fits-all,” and are only effective for 60% of patients. The current study aims to use advanced statistical methods and a novel theoretical framework to map the stability of tic patterns, along with systemic factors that relate to tic change over time. Study hypotheses, based on the literature and preliminary data, are that a) tic change patterns will be stable before intervention for all participants, b) disruption of stable patterns during the intervention phase will be associated with treatment response, and c) this disruption will depend on the specific driver of tic symptoms pre-intervention. N = 30 youth ages 12-17 with chronic tics will be recruited for the study. There will be three study phases: 1) pre-intervention (4 weeks), 2) intervention (8 weeks), and 3) post-intervention (4 weeks). Before and between each phase, participants will complete 4 traditional assessments to assess symptoms and treatment response. Throughout the 16 weeks of the study, we will collect EMA data focused on factors relevant to tics (4x per day), physiological data from wearable devices (passive, continuous), and neurocognitive task performance and tic video observation (1x per week). Results will inform efforts to develop individualized interventions for individuals with PTDs to improve treatment outcomes.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Duchenne muscular dystrophy (DMD) is one of the most severe neuromuscular diseases, with a median life expectancy of 22 years. DMD results from the absence of the protein dystrophin, reducing the ability of muscle to respond to stresses imposed by force generation and resulting in significant contractile dysfunction. In human patients with DMD, this manifests in progressive skeletal muscle weakness beginning around 3-5 years of age and cardiomyopathy and respiratory insufficiency around age 25, leading to death. In the mdx mouse model of DMD, the absence of dystrophin also results in rapid and robust deficits in contractility, particularly loss of force with repetitive eccentric contractions (ECCs) in skeletal muscle. The mechanisms by which mdx mice undergo ECC force loss, and those which result in broader contractile deficits of striated muscle, are poorly understood. Preliminary data from the Ervasti lab implicates elevated reactive oxygen species (ROS) and subsequent hyperoxidation of the thiol proteome in DMD pathophysiology. Moreover, supplementation with the gasotransmitter hydrogen sulfide (H2S), which is reduced in mdx muscle, completely protects mdx extensor digitorum longus (EDL) muscle from ECC force loss in vitro. The mechanisms by which H2S protects from ECC force loss, and its contributions to contractility in general, are not well characterized in the context of dystrophin- deficient muscular dystrophy. The studies I designed in my proposal will test the hypothesis that elevations in H2S will increase the persulfidation of reactive thiol residues on cysteines of contractile proteins, thereby preventing hyperoxidation and associated contractile dysfunction. The experiments proposed here will strive to characterize the effect of chronic, systemic H2S supplementation on DMD pathophysiology in vivo (Aim 1) and define the effects of low H2S levels on oxidative post-translational modifications in mdx contractile proteins (Aim 2). A more complete understanding of contractile dysfunction and protection from force loss in mdx muscle is critical to generating more efficacious therapeutics. In addition to the research proposal, the following documents include a comprehensive and rigorous set of research and clinical training plans that will allow for the successful completion of my MD and PhD degrees and will enhance my development into a productive physician-scientist, able to drive cutting-edge translational research and clinical care for my patients.

NIH Research Projects · FY 2025 · 2024-08

ABSTRACT Natural killer (NK) cells are key mediators of anti-tumor immune responses, acting to directly kill cancer cells (CD56dim subtype) and recruit pro-inflammatory immune populations (CD56bright subtype). Recent discoveries, however, have revealed other NK cell subtypes with properties of tissue-resident and adaptive, memory-like phenotypes. Tissue-resident NK cells have been shown to have elevated production of chemokines, such as CCL3 and XCL1 that, in turn, recruit conventional type 1 dendritic cells (cDC1) and CD8+ T cells to tumors. Adaptive NK cells have been shown to persist for months longer than conventional NK cells and display enhanced tumor cell killing. Given the diversity of the NK cell repertoire, it is crucial to identify which NK cell subtypes exist in tumors and are key to anti-tumor responses. To address this, our group has generated a pan- cancer atlas of NK cells using single-cell RNA-sequencing data from 326 patients across 17 cancer types. In doing so, we identified prominent populations of CD56bright NK cells that display gene signatures suggestive of both tissue-resident and adaptive NK phenotypes (bright-taNKs). We found bright-taNKs to display higher expression of chemokines and cytokines, such as CCL3, XCL1, and IFNG, suggesting a functional role in immune cell recruitment to tumors. Using a gene signature to estimate the relative abundance of bright-taNKs in bulk RNA-seq data, we found that bright-taNKs are associated with significantly longer disease-free survival in cancer patients. Thus, bright-taNKs may be critical for anti-tumor immunity and represent attractive phenotypes to mimic for the next generation of engineered NK cell therapies. As little is currently known about bright-taNKs, this proposal aims to advance our understanding of the functions of bright-taNKs in tumors and identify mechanisms promoting their distinct phenotypes. In Aim 1, we will use orthogonal methods to determine whether bright-taNK populations are associated with increased recruitment of anti-tumor immune populations, such as cDC1s and CD8+ T cells. We will develop a bioinformatics tool that can be paired with well-established tools to estimate fractions of different NK cell subtypes and other immune cell types in bulk RNA-seq data. Additionally, we will apply highly-multiplexed spatial imaging tools to interrogate the spatial distribution of bright- taNKs, and other NK cell populations, in tumors and their associations with other immune cell types. In Aim 2, we will use gene delivery platforms to screen for transcription factors that promote features of tissue-resident and adaptive phenotypes in CD56bright NK cells. We will then transition the most promising gene targets to our clinical-grade induced pluripotent stem cell to NK cell (iNK) platform to holistically evaluate the effects on anti- tumor functions. Altogether, this proposal aims to advance our understanding of bright-taNKs, and other NK cell subtypes, in tumors. Ultimately, drivers of favorable NK cell properties can be readily integrated into our group’s bench-to-bedside pipeline for NK cell therapies to enhance their efficacy and durability.

NIH Research Projects · FY 2025 · 2024-08

ABSTRACT Child and adolescent behavioral health problems are related to the leading causes of youth morbidity and mortality and are costly to society. Parent-focused interventions effectively prevent behavioral health problems such as depression and conduct disorders and can provide an economic benefit to society. Unfortunately, parenting programs are not widely available, not accessible, nor well-attended. Pediatric primary care is a non- stigmatizing setting with nearly universal reach and, therefore, an ideal access point to increase access. However, primary care clinicians (PCCs) often have insufficient training in behavioral health topics and typical referral practices are often inadequate. There are also logistical barriers to attending in-person parenting programs, like the need for childcare and a large time-commitment. There is a need to develop effective referral practices in conjunction with increasing the accessibility of parenting programs. Our long-term goal is to prevent significant behavioral health problems through widespread access to effective and accessible parenting programs through primary care referrals. Previously, we successfully developed and pilot tested a brief training for PCCs that we call Support & Guide (S&G). S&G training provides a communication strategy for engaging parents in a conversation about parenting and for making referrals. We also successfully pilot tested a brief, online parenting intervention based on GenerationPMTO, called Empowered Generations (eGen); eGen is a six-session parent training program provided by therapists via video chat online to parents with a child between the ages of 3-8 years. S&G training and eGen were both found to be feasible and acceptable, and both demonstrated promise of effectiveness. Significantly, all clinical aspects of the study have been conducted in real-world settings and systems and are highly pragmatic. In the proposed R01, we will build on the knowledge gained through the R34 to conduct fully powered randomized trials. The secondary objective of this study is to test the effectiveness of brief PCC S&G training. The primary objective of this project is to test the effectiveness and cost-benefit of eGen. The study design includes two parallel randomized controlled trials (RCT). In the S&G RCT, we will randomize PCCs (aim 1); in the eGen RCT we will randomize parents (aim 2). These two trials are linked by the fact that the parents referred by PCCs in aim 1 are the same parents who are invited to participate in the aim 2 eGen trial. In addition, we will examine the cost benefit of eGen (aim 3) when implemented with community therapists and with a real-world, primary-care-referred sample of parents. This research is significant because it simultaneously addresses the need to develop effective referral practices in conjunction with increasing the accessibility of parenting programs. The innovative design in this study captures the entire patient flow from initial engagement to outcomes and operates within real-world systems and documents the potential return on investment for payors and society.

NIH Research Projects · FY 2024 · 2024-08

Biomedical HIV prevention has extended beyond pill-based strategies to now include long-acting injectable HIV pre-exposure prophylaxis (LAI-PrEP) dosing regimens, yet implementation of LAI-PrEP is suboptimal to date. LAI-PrEP has unique implications for clinical practice that will not easily fit within implementation strategies for oral PrEP. A better understanding of how these specific factors impact roll-out is needed to address enduring barriers and guide implementation considerations. As such, this proposal aims to: (1) identify enduring barriers and facilitators to LAI-PrEP implementation in the U.S., (2) determine optimal implementation strategies for LAI-PrEP in partnership with stakeholders, and (3) pilot an implementation strategy within two outpatient clinics in the Midwest. To help guide this formative work, the team leverages the Consolidated Framework for Implementation Research to understand determinants of LAI-PrEP implementation and uses two Expert Recommendations for Implementing Change strategies rated with high feasibility and high importance to maximize potential impact in supporting LAI-PrEP roll-out. Preliminary impact of LAI-PrEP implementation will be evaluated using RE-AIM, assessing Reach, Effectiveness, Adoption, Implementation, and Maintenance via electronic medical records review, survey, and exit interviews. Exit interviews will also determine feasibility, acceptability, and appropriateness of the implementation strategy. This projects’ target on LAI-PrEP is timely, uses an innovative sampling approach to study providers’ perspectives, and is positioned to bring forth paradigm-shifting approaches to incorporate LAI-PrEP into clinical practice. This proposal is led by an experienced early-stage investigator in collaboration with a multidisciplinary investigative team, compliments research supporting oral PrEP implementation, and sets the stage for larger scale implementation efforts to enhance integration of LAI-PrEP into clinical practice.

NIH Research Projects · FY 2025 · 2024-08

Project summary/abstract. The emerging field of allosteric inhibition of protein kinases has the potential to provide highly targeted therapeutics for diverse diseases without undesirable side effects. Our long-term goal is the discovery and development of highly selective allosteric inhibitors for side effect-free cancer chemotherapy and for reversible non-hormonal contraception. The overall objective of this application is to identify novel allosteric inhibitors for four kinases: WEE1-like protein kinase 2 (WEE2), testis-specific serine/threonine kinase 1 (TSSK1), cyclin-depended kinase 11 (CDK11), and cyclin dependent kinase 2 (CDK2) by screening DNA encoded libraries (DEL) consisting of 2B compounds. The four kinases are validated drug targets: WEE2 for female non-hormonal contraception, TSSK1 for male non-hormonal contraception, CDK11 for cancer chemotherapy and CDK2 for cancer and male and female non-hormonal contraception. WEE2, TSSK1 and CDK11 are on the NIH list of underexplored targets. CDK2 inhibitors have been studied for cancer application but limited studies have been carried out for male contraception and none for female contraception. Hit compounds from the DEL screen will be resynthesized by conventional synthesis and then confirmed by using functional biochemical assays, orthogonal confirmatory assays, and biochemical and biophysical assays to determine allosteric binding. Validated compounds will be tested in a 24 kinome screen for kinase selectivity and in a limited set of in vitro ADMET assays to assess their drug-like properties. CDK11 and CDK2 inhibitors will be tested in cancer cell lines that express their respective targets. TSSK1 inhibitors will be tested using a computer-assisted sperm analyzer (CASA) for sperm motility measurements. Selective allosteric inhibitors identified in this project are expected to be useful probes to investigate the biology of the four kinases and to serve as early leads for drug discovery.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ABSTRACT The use of specific pathogen free (SPF) mice has been instrumental in the advancement of immunological research, reducing variability between experiments and institutions. However, it is becoming increasingly evident that pathogen experience alters the immune system dramatically. This limits the translation potential of discoveries in SPF mice, as humans are regularly exposed to pathogens from birth. Work from the Hamilton lab and others has found that increasing the microbial exposure of mice through pet shop mouse cohousing (COH) alters the immune compartment of mice to more accurately reflect that of an adult human. This change is particularly evident in the memory CD8 T cell compartment. The CD8 T cell memory pool contains cells with a wide range of trafficking patterns, effector functions, and longevity. In SPF mice, the CD8 T cell compartment is dominated by naïve cells and central memory T cells. However, COH mice and adult humans contain predominantly effector memory T cells, including long-lived effector cells (LLEC), a population described by the Hamilton Lab. This proposal aims to understand how pathogen exposure and inflammation lead to the expansion and maintenance of LLEC, and how the expansion of this cell population alters the overall function of the T cell compartment. Aim 1 will determine what proinflammatory cytokines support LLEC formation and persistence. Experiments will identify cytokines and other factors that lead to the increase in LLEC during COH, via the use of RNA sequencing and CRISPR mediated genetic knockouts. We will also determine if we can expand LLEC in the absence of pathogen exposure via administration of cytokines. Aim 2 will determine how COH alters the function of LLEC, and address the consequences of skewing the memory compartment to LLEC for responses to systemic and local infections. This aim will utilize adoptive cell transfers and an inducible fluorescent reporter to determine if COH LLEC can protect against systemic and local infections. Focusing on CD8 T cell memory, and incorporating RNA sequencing analysis, will support goals 1 and 2 of my training plan, and the proposal will develop my skills as an academic scientist as a whole (training aim 3). The Center for Immunology at the University of Minnesota is an ideal environment to complete this proposal as there is a strong record of impactful immunological research and postdoctoral training, particularly within T cell biology. Altogether, these studies will yield critical information regarding the importance of pathogen exposure and inflammation on the memory T cell compartment, as well as novel information on the functionality of subsets of memory CD8 T cells. These data will be useful to develop vaccines and T cell therapies with increased translation potential.

NIH Research Projects · FY 2026 · 2024-08

The rural U.S. is sick, old, and in decline – or is it? This familiar narrative of “left behind” rural people and places tells only one part of the story of rural America – the most pessimistic one. Although it is true that rural areas are home to disproportionate shares of older and sicker people, and rural-urban disparities in health and longevity are large and growing, it is also true that some rural places are thriving. New approaches to researching rural health and aging are needed, recognizing that rural America is not monolithic; it contains both resilient and vulnerable places, and its challenges are multilevel and multidimensional. Building on the successes and lessons of its last five years, the Interdisciplinary Network on Rural Population Health and Aging (INRPHA) will spark and sustain new collaborations that will advance research on the factors affecting the health and wellbeing of rural working-age and older adults within the context of prevailing demographic trends, slow-moving macro-level stressors, and contemporary public health and environmental shocks. In so doing, the Network will inform new approaches to improve wellbeing, health, and functioning in rural America. Our proposal is both significant and novel because INRPHA’s activities will a) be national in scope and attend to differences in health and aging across different rural regions, economies, population change patterns, and demographic groups; b) illuminate not only rural-urban, but also within-rural variability to better understand variation in outcomes, including what we can learn from thriving rural communities; c) elucidate mechanisms across the life course that are driving observed rural-urban and within-rural health and aging outcomes, trends, and disparities; and, d) leverage ex- isting NIH-funded data resources that can be used to advance rural population health and aging research. Lev- eraging the institutional assets within the proposed Network’s five lead universities – U Minnesota, Penn State, Syracuse U, Mississippi State, and U Colorado, Boulder – INRPHA’s aims are to (1) conduct and support re- search that will enhance understanding of the multilevel and multidimensional causes of health, functioning, and mortality outcomes, trends, and disparities among U.S. rural working-age and older adults, with a particular focus on within-rural heterogeneity; (2) strategically grow the Network by expanding existing membership to include scholars from relevant disciplines and who represent various topical and methodological perspectives; (3) pro- vide development and training opportunities through pilot grants, structured mentoring, topical and data work- shops, and regular network meetings, including efforts to promote members’ knowledge and use of existing NIH- funded datasets conducive to conducting robust rural-urban and within-rural research; and, (4) advance mem- bers’ dissemination of research findings through training and support of academic articles and policy briefs and through public webinars to expand access to knowledge that will inform research, policies, and programs related to rural population health and aging. INRPHA’s activities emphasize developing a sustainable foundation to sup- port continued innovative, publicly accessible, and impactful research on rural population health and aging.

NIH Research Projects · FY 2025 · 2024-08

PROJECT ABSTRACT Cognitive changes occur as a normal process of aging and among the most important changes in cognition in normal aging are declines in cognitive flexibility (CF). CF is considered a core aspect of executive functioning and describes an individual’s ability to adapt their behavior and thinking to a changing environment. Disruptions in CF impact functional independence, communication with others, and socialization. Despite the well-documented prevalence of CF decline in aging, and its exacerbation in neurodegeneration, the mechanisms that lead to structural and functional changes involved in the regulation of CF are not well understood. The majority of studies related to CF impairment have focused on changes within the hippocampus and cortex, however, the dorsal striatum, particularly the dorsomedial striatum (DMS), has long been strongly associated with CF. The dorsal striatum is traditionally associated with control of motor function, but recent human fMRI and PET analyses demonstrated that declines in striatal function predict declining cognitive abilities during aging. Striatal function is governed by the innervation of two excitatory glutamatergic circuits: cortico- (C-S) and thalamo-striatal (T-S). Importantly, T-S synapse density preferentially declines during aging and is associated with declining CF performance. However, the regulatory mechanisms by which T-S synapses are preferentially lost in the aging striatum and how they contribute to CF decline is unknown. Depletion of the heat shock transcription factor 1 (HSF1), a transcription factor canonically known for its role in cellular stress responses, within the striatum, accelerates T-S synapse loss and results in impairment of CF. HSF1 levels also decrease in aging and HSF1 has been shown to directly regulate the transcription of synaptic genes within the striatum and other brain regions. Taken together, the goal of this proposal is: 1) to determine the molecular underpinning that govern the differential effects of HSF1-mediated synaptic regulation within the DMS and 2) to understand the role of HSF1 and T-S synapses in striatal dysfunction and CF impairment. I will fill these gaps in knowledge through two complementary aims. In Aim 1, I will examine the role of HSF1 in the regulation of the synaptoproteome of excitatory striatal projections through proteomic analyses of T-S and C-S synaptic terminals and synapses from a conditional HSF1 knock-out mouse. In Aim 2, I will assess the contribution of striatal HSF1 to CF by deleting HSF1 within the adult DMS using AAVs expressing Cre- recombinase and the necessity of T-S connectivity to CF by optostimulation of T-S terminals in DMS. Successful completion of this proposal will provide a mechanistic understanding of how synaptic dysfunction within the DMS contributes to CF which has implications in aging and neurodegeneration, as well as establish the role of HSF1 in the regulation of striatal glutamatergic synapses and CF that may result in new therapeutic strategies to prevent CF deficits in the aging population.

NIH Research Projects · FY 2026 · 2024-07

Project Summary/Abstract The deltaretroviruses represent one of seven genera in the Retroviridae and are recognized as being complex retroviruses that encode for gag, pol and env along with several important accessory genes. These viruses have been associated with the development of lymphoid leukemias in humans (i.e., human T-cell leukemia virus, HTLV) and cattle (i.e., bovine leukemia virus, BLV). Deltaretroviruses are notorious for being difficult to propagate in cell culture, which has prohibited rigorous analyses of virus replication, including the steps involved in retrovirus assembly. The detailed steps involved in retrovirus assembly are poorly understood even for the more tractable systems, including human immunodeficiency virus (HIV). Our preliminary studies have revealed that the deltaretroviruses have distinct immature and mature particle morphologies compared to other retroviruses, as well as distinct Gag-Gag and capsid-capsid (CA-CA) lattice structures, which likely impacts virus particle formation. Furthermore, we have made observations which reveal that HTLV Gag puncta biogenesis is distinct among retroviruses, relying heavily on Gag recruitment to particle assembly sites by Gag already at the plasma membrane. This distinctive pathway for particle assembly likely has important implications for establishment of HTLV particle assembly sites and the dynamics of particle assembly. In this application, we propose to advance our research in novel directions through innovative state-of-the-art experimental approaches, making use of comparative retrovirology in order to provide deep insights into the nature of deltaretrovirus particle assembly and maturation. In particular, we will apply cryo-electron microscopy/tomography (cryo-EM/ET), super-resolution microscopy, total internal reflection fluorescence (TIRF) microscopy, and dual-color z-scan microscopy to investigate the 1) structure-function analysis of deltaretrovirus maturation, 2) deltaretrovirus Gag recruitment and puncta biogenesis, and 3) analysis of critical steps in deltaretrovirus particle biogenesis. The proposed studies will extend our preliminary studies and exploit advanced imaging technologies to gain new insights into the deltaretrovirus particle assembly pathway, which we have demonstrated as being distinct from other retroviruses. This research will provide foundationally important knowledge regarding the details of virus assembly at the plasma membrane for enveloped viruses.

NIH Research Projects · FY 2025 · 2024-07

ABSTRACT: Alpha-synuclein (aSyn) accumulation and misfolding is implicated in the pathogenesis of Parkinson's Disease (PD) and related synucleinopathies. These disorders include multiple cellular dysfunctions including impaired proteostasis (e.g., the dysregulation of the autophagy lysosomal pathway [ALP]). As preclinical disease models become more complex to better recapitulate disease relevant pathophysiology— i.e., monoculture ➔ 2D co- culture ➔ 3D organoids (both mono- and co-cultured) ➔ in vivo — advanced multiplexing technologies that are capable of dynamic temporal and spatial monitoring of protein-protein interactions (e.g., aSyn oligomerization and misfolding) and pathological phenotypes are required. Existing fluorescence-based biosensors are limited by their static nature and inability to differentiate signals across distinct cell types within multi-cellular environments. To address this, our proposal introduces Dark-FRET (DF) and Dual-Channel DF (DCDF) cellular biosensors that utilize the Shadow-G/Y/R series of acceptor protein which were engineered as to have reduced quantum yield and negligible fluorescence emission, eliminating emission spillover from acceptor proteins which facilitates improved live-cell multiplexing for multiple protein-protein interaction (PPI) fluorescence assays. This cutting-edge approach enables real-time monitoring of both aSyn folding (ShadowY-aSyn-mNeongreen) and aggregation (aSyn-mScarlet-I3/ShadowR) FRET biosensors expressed in distinct cellular populations. We expand on these capabilities by multiplexing the aSyn DCDF biosensors with our TFEB (the master regulator of ALP), FRET and nuclear translocation biosensors. Aim 1 involves applying these biosensors to CNS-resident cell lines (neurons, microglia, astrocytes) to establish mono-, bi-, and tri-culture cellular models for dynamically tracking cell-specific aSyn interactions and ALP phenotype. Aim 2 extends DCDF technology to in vivo models using novel C. elegans strains expressing aSyn DCDF and ALP biosensors in specific tissues. By enabling simultaneous monitoring of aSyn oligomerization and phenotypic dysfunction, this approach provides a valuable tool for investigating synucleinopathies. Its potential applications extend to 3D organoids and hiPSC models, offering insights into diverse cell-type pathways and enabling the identification of compounds modulating aSyn oligomerization and pathological phenotypes in these advanced preclinical models. In summary, our DF/DCDF technology presents a groundbreaking approach, enabling dynamic monitoring of aSyn aggregation and pathological phenotypes across complex disease models, paving the way to address biological questions on the supporting roles of microglia and astrocytes on neuronal health and function. Furthermore, these multiplexed biosensors represent an innovative preclinical therapeutic discovery platform that holds promise for enhancing the drug discovery process, potentially filling the therapeutic gap in synucleinopathy treatment.

NIH Research Projects · FY 2025 · 2024-07

Project Summary/Abstract Generation of a functional adaptive immune response relies on the collaboration of the cellular (T cell) and humoral (B cell) compartments. T follicular helper (Tfh) cells provide essential help to B cells to undergo isotype class-switch recombination and generate high-affinity antibody through somatic hypermutation. Follicular regulatory T (Tfr) cells have been implicated in controlling this process as they have been shown to be critical in regulating germinal center B cell responses and prevent autoantibody formation. However, how these Tfr cells develop is unknown. B cells undergo a unique type-III interferon dependent and isotype class- switching process in the thymus, the location of T cell development, and induce clonal deletion and Treg cell selection. Developing Treg cells that depend on licensed thymic B cell antigens may also interact with activated B cells in the periphery and take on a Tfr phenotype. Thus, Treg cells selected by thymic B cells may become Tfr cells in the periphery and help to reduce the risk autoantibody generation. The objective of this work is to understand what role licensed thymic B cells play in T cell tolerance. In this proposal we identified T cell receptors that generate Tfr cells when expressed by developing T cells, propose to study the requirements of these TCRS for selection as well as explore further functional consequences when thymic B cells are absent. Our central hypothesis is that type-III interferon drives thymic B cell licensing, resulting in the presentation of B cell activation induced self-peptides, thereby supporting the development of Treg cells that become Tfr cells and regulate humoral immune responses. Through this work we hope to generate a mechanistic understanding of the impact of thymic B cell activation on immune tolerance. These findings will have the potential to reveal new pathways regulating adaptive immune responses and have implications in our understanding of autoimmune diseases like systemic lupus erythematous or rheumatoid arthritis, diseases where pathogenic autoreactive antibodies mediate disease. I am applying for this K99/R00 as an Instructor in Laboratory Medicine and Pathology Department with subspecialty training in Molecular Pathology. My long-term career goals are to support for the clinical diagnosis of autoimmunity and inborn errors of immunity and to lead an R01 funded research program that investigates central tolerance and immunodeficiency.

NIH Research Projects · FY 2026 · 2024-07

Summary The thymus plays a critical role in preventing autoimmunity by deleting or inducing a Treg cell fate in thymocytes that react to self-antigens present in the thymus. But it is unclear how the thymus achieves tolerance to transiently expressed self-antigens, like those expressed in induced immune states such as inflammation, wound repair, and germinal center reactions. We have recently discovered that the thymus constitutively produces type I and III interferons and class switched activated B cells, beginning quite early in life. Our published and preliminary data show that these cytokines activate antigen presenting cells in the thymus environment and cause reproducible changes in the T cell repertoire selected, particularly the Treg cell repertoire. Thus, we seek to test the hypothesis that T cells need to be highly tolerant of induced immune states (inflammation, germinal centers) in order to function in those states without inducing autoimmune pathology. The proposed research will: 1) Test if thymic IFN provides T cell tolerance to interferon stimulated self-antigens, 2) Determine why IFNI/III deficient mice have a major thymic selection defect, and 3) Test if IFNIII-licensed B cells select Treg cells that regulate humoral immune responses. These aims will provide a mechanistic understanding of the source and consequences of innate immune activation in the thymus. Understanding T cell tolerance to self-antigens in immune activated states is important because many human autoimmune diseases are associated with or follow infections.

NIH Research Projects · FY 2026 · 2024-07

Project Summary The highly conserved Planar Cell Polarity (PCP) pathway is essential for cells to communicate directional information between neighboring cells to drive collective cell movements and oriented cell behaviors during embryonic development. Disruptions in PCP signaling lead to severe developmental disorders including congenital heart defects, neural tube closure defects, and shortened body axes. The core PCP proteins are transmembrane proteins that asymmetrically polarize to opposite sides of the cell. Extracellularly, the proteins form an asymmetric bridge that directly couples the polarity of one cell to its adjacent neighbors. In this way, the PCP proteins self-organize to coordinate cell polarity at the local level. A hallmark of PCP polarity, however, is its tissue-wide coordination, spanning hundreds or thousands of cells across great distances. The self-organizing properties of the core pathway alone cannot account for this long-range coordination as self-organization could occur spontaneously in any direction. Further, modeling has shown that self-organization can only propagate locally, resulting in a swirling pattern of PCP polarity rather than tissue-level coordination. For this reason, the PCP field has come to a consensus that ‘directional cues’ are key missing factors that must link the PCP axis to the tissue axis. Directional cues are thought to act in a gradient across a tissue where they bias the distribution of the PCP proteins along the same axis by regulating their trafficking, stability, or turnover. Despite a general consensus on how directional cues could pattern PCP organization, the identities of these cues remain elusive. As such, the mechanism of their action cannot be interrogated. A major challenge in identifying directional cues is that they are also required for tissue growth and differentiation. Thus, the tissue of interest becomes compromised in the absence of the cue, making it difficult to investigate the cue’s role in organizing PCP in isolation. Epidermal tissues provide a striking readout of planar polarity in their uniform alignment of bristles, scales, fur, and feathers across an animal’s body. Remarkably, spontaneous mutations in small mammals and birds have produced animals with region-specific misorientation of epidermal polarity. From these phenotypes, we hypothesize that the skin is compartmentalized into region-specific domains that require multiple directional cues to coordinate polarity across the tissue. Further, the fact that the skin is morphologically normal under these conditions despite the polarity defects demonstrates that the skin can be leveraged as a model system to interrogate directional cues. Our long-term goal is to identify and study the directional cues that pattern PCP across the mouse epidermis to reveal mechanisms that coordinate long-range polarity. By using natural variants and taking a candidate approach, we will identify and characterize directional cues and will reveal how they alter the dynamics of PCP asymmetry along a tissue axis. This approach addresses critical and long-standing questions in the PCP field and will shed light on fundamental mechanisms that pattern embryonic tissues.

NIH Research Projects · FY 2025 · 2024-07

The mission of the Clinical and Translational Science Institute (CTSI) postdoctoral T32 Program is to contribute to the nation’s biomedical workforce by training an interdisciplinary pool of scientists in clinical and translational science (CTS) and leadership. We will meet this mission by developing, implementing, and continually improving a training program for postdoctoral trainees (Scholars) that integrates a mentored research experience with an individualized curriculum combined with professional development activities highlighting our institutional strengths in community engagement, data science, team science and effective communication skills. The UMN CTSI T32 Program is built upon our current successful Translational Research and Career Development (TRACT) TL1 Program, which guided the development of early-stage CTS researchers over the past 4 years. Through personalized mentorship, coursework, seminars, and other tools, the TRACT program has guided 6 postdoctoral Scholars along a rigorous path of professional development in CTS. Moving forward, the new UMN CTSI T32 Program’s mission is to educate and train Scholars from across the UMN system in the fundamentals of CTS through a 2-year comprehensive yet individualized program. We will recruit and train 3 cohorts of 4 postdoctoral Scholars from across a variety of disciplines at the UMN. UMN CTSI T32 will provide a research experience with mentors trained in evidence-based mentoring, focusing on collaboration and communication with community members throughout the research process, inclusive of a Community Mentor and team science across the UMN ecosystem. The UMN CTSI T32 will also include near-peer learning experiences with other CTSI program Scholars as well as provide education and opportunities for a variety of career paths. The UMN CTSI T32 will use a comprehensive and adaptive evaluation plan for rigorous programmatic assessment and continuous quality improvement to maximize the training experience. The principal Scholar outcome is to have a deep knowledge in the science of translation beyond translational research. Another key Scholar outcome is an understanding of the importance of, and key strategies for, building truly collaborative interdisciplinary teams composed of people from a variety of scientific backgrounds whose collective roles synergistically optimize the advancement of diagnostics, therapeutics, and clinical interventions that improve health. Other outcome measures include the ability to communicate with other scientists and nonscientific audiences and collaborators through our intensive formalized training in communicating science, and the ability to effectively engage with communities through intensive community engagement training including a Community Mentor Program. Programmatic outcomes include the development of training modules and workshops on the most effective community engagement and communications strategies exportable to the national CTSA consortium.

NIH Research Projects · FY 2024 · 2024-07

PROJECT SUMMARY Abstract The refinement of gross motor skills, such as locomotion, during development is conserved across vertebrate species. As an example, locomotor output produced by immature animals is initially coarse and becomes progressively more refined during development. Although we previously demonstrated, in larval zebrafish, that neuromodulatory signaling via the dopaminergic system was necessary for the developmental transformation of locomotor (swimming) activity from an immature to a mature state, the underlying molecular and functional mechanisms remain elusive. Our goal here is to characterize the functional and molecular mechanisms underlying the dopaminergic-mediated refinement of spinal locomotor activity using an established vertebrate model system (larval zebrafish) and a well-validated complement of tools, including electrophysiology, pharmacology, in vivo functional imaging (calcium and dopamine), and quantitative reverse transcription polymerase chain reaction (qRT-PCR). The results of this project will serve as the basis for a future R01 proposal that will examine the causal relationship between the developmental changes in the dopaminergic system and locomotor refinement. Overall, since the dopaminergic and motor systems are highly conserved across vertebrates, we expect these findings to translate to other animals, including humans.

NIH Research Projects · FY 2026 · 2024-07

PROJECT SUMMARY/ABSTRACT Over 75% of people with Parkinson's disease (PD) have significant sleep-wake disturbances that are major contributors to decreased quality of life and can be more disabling and resistant to treatment than the motor symptoms of PD. Currently, the mechanisms contributing to disordered sleep in people with PD are poorly understood and there is a critical need for therapeutic inventions to improve sleep quality. This project will provide new insight into the pathophysiology of sleep-wake disturbances in PD by characterizing the changes in sleep- related neuronal activity and physiological interactions that occur between subcortical and cortical structures in the basal ganglia thalamocortical (BGTC) circuit during progressively more severe parkinsonian states. We will expand our previous work to include the pedunculopontine nucleus (PPN) given its critical role regulating sleep- wake states and extensive connectivity to the BGTC, exploring the changes in coupling and connectivity that occur within and across the BGTC-PPN network that underlie disordered sleep in PD secondary to dopaminergic loss in the substantia nigra (Aim 1). It will compare how deep brain stimulation (DBS) in the STN, GPi, and GPe modifies subcortical-cortical interactions in the BGTC-PPN circuit to influence sleep-wake behavior and elucidate the fiber pathway activations underlying these changes (Aims 2 and 3). This study will provide data with immediate translational value by identifying whether DBS in one target is more effective than another in normalizing sleep-related neuronal activity and improving sleep-wake behavior. Furthermore, knowledge about how changes in neuronal activity across the BGTC-PPN network correlates with altered sleep from normal, parkinsonian, and parkinsonian+DBS conditions will provide the basis to develop more effective stimulation strategies that utilize target-specific physiological biomarkers and closed-loop DBS control paradigms tailored to individual patient's sleep disturbances. These data will also provide the basis to target specific pathways with DBS to optimize sleep-related outcomes in PD.

NIH Research Projects · FY 2026 · 2024-07

PROJECT SUMARY / ABSTRACT In general, enzymes are very precise at catalyzing a specific canonical reaction that fits within a particular metabolic network. Still, no enzyme is a perfect catalyst. The inherent flexibility of proteins makes it difficult for enzymes to distinguish their canonical substrate from structurally related compounds. Thus, many enzymes act on unintended substrates (i.e., substrate promiscuity). Substrate promiscuities result in the formation of unintended or damaged metabolites (i.e., metabolite damage) that can be a useless drain on metabolism, and may be inhibitory and/or reactive, sometimes leading to toxicity. Accordingly, metabolite damage repair enzymes exist for the specific purpose of counteracting metabolite damage, often by converting a damaged metabolite to a canonical one. The physiological importance of metabolite damage and its repair has been revealed over the past ~15 years as a handful of metabolic diseases in humans were discovered to be caused by disruption of metabolite damage repair genes, many of which are highly conserved across the three domains of life. The proposed project will address metabolite damage repair associated to the TCA cycle – a universal core metabolic pathway that is involved in energy conversion and is a source of chemical building blocks that supplies much of metabolism. The TCA cycle is a hotspot for metabolite damage due to high carbon flux through the pathway and chemical intermediates that are structurally similar organic acids, which can engage in promiscuous side reactions catalyzed by the abundant cycle enzymes. I have identified and characterized several highly conserved metabolite damage control systems related to vitamin, cofactor, and amino acid metabolism, and my training has empowered me with a unique skillset and perspective that is allowing me to make similar discoveries related to the TCA cycle. One enzyme that I have identified is particularly intriguing. A prevalent side-reaction of the TCA cycle enzyme succinate dehydrogenase oxidizes malate to enol-oxaloacetate (OAA), a metabolically inactive form of OAA that is a potent inhibitor of the TCA cycle. Our results provide strong evidence this side reaction is one of the most prevalent promiscuous reactions in nature, and that enol-OAA is a potent inhibitor of the TCA cycle. We identified a universally conserved enzyme, OAT1, that removes the inhibitor, and show that bacterial cells lacking OAT1 have a severely attenuated TCA cycle. The proposed work will integrate biochemical, genetic, and metabolomics/ fluxomics approaches to determine how OAT1 impacts the physiological and metabolic states of prokaryotic and single and multicellular eukaryotic model organisms. Completing this project will lead to the detailed characterization of a previously unrecognized but critical aspect of the TCA cycle, ultimately redefining one of the most universal core metabolic pathways in biology. This work will also provide insights into mitochondrial metabolism and physiology that will impact human health and disease, and deliver a metabolite damage repair enzyme for use in optimizing synthetic biology platforms.

NIH Research Projects · FY 2025 · 2024-07

Project Summary Tick-borne Powassan virus (POWV) is an emerging public health threat, but has been understudied to date. Since its discovery in 1958, human cases of POWV have been reported in the United States, Canada, and Russia. Starting in 2006, confirmed cases of POWV have increased in both frequency and scope in New England and the Upper Midwest. Notably, POWV is now considered an emerging public health threat because it has been frequently associated with the aggressive, human-biting blacklegged tick vector, Ixodes scapu- laris. The future spread of POWV is unpredictable, but it will be important to understand the potential for more widespread emergence. The goals of this project are to evaluate whether there are regional, local, and even hyperlocal differences in the genetic structure of tick and POWV populations that affect patterns of POWV host/vector competence, virulence, and evolution. These questions are important because our knowledge of how tick-borne flaviviruses are maintained in distinct transmission foci and how new transmission foci emerge is limited. Accordingly, through this NIH/NIAID R01, we will seek to examine these relationships across Minne- sota and New York, two states with increasing burdens of tick-borne disease risk. There are 3 specific aims: 1. Assess the distribution and genetic structure of POWV and Ixodes scapularis from distinct foci in the Northeast (NY) and Midwest (MN) U.S. 2. Evaluate the extent to which tick population interacts with viral genotype to drive local adapta- tion and regional variability in Ixodes scapularis competence for POWV. 3. Determine how POWV genetic variability and regional selective pressures impact host compe- tence and disease heterogeneity. In these studies, we will expand POWV surveillance activities in MN to intensely sample POWV genomes and tick populations from across the state. The NY State Department of Health has a robust POWV surveillance program in place for this project that we will leverage. We will also seek to understand the phenotypic and ge- notypic underpinnings of virus–host dynamics for POWV using mouse and tick experimental systems. These studies are critically important because POWV likely will continue to emerge in the future. Therefore, under- standing how POWV overcomes evolutionary barriers to emerge and cause disease in humans will be critical for prediction, prevention, and control of this arboviral disease.

NIH Research Projects · FY 2025 · 2024-07

ABSTRACT The goal to effectively treat neurodegenerative disorders requires an understanding not only of neuronal dysfunction, but also of dysfunction of non-neuronal cells that can initiate and contribute to neuronal pathology. Spinocerebellar ataxia type 1 (SCA1) is a fatal, dominantly inherited neurodegenerative disease caused by the abnormal expansion of CAG repeats in the Ataxin 1 (ATXN1) gene. SCA1 patients suffer from progressive neuronal degeneration and reactive astrogliosis, especially in the cerebellum, leading to motor deficits. Despite intense research focus on the disease mechanisms in cerebellar neurons, there are no effective therapies available to cure, delay or ameliorate SCA1. Astrocytes are brain cells that play fundamental roles in nearly all aspects of neuronal and brain function. Recently, using single-nuclei RNA sequencing we demonstrated that cerebellar astrocytes express Atxn1 at a level comparable to neurons (Borgenheimer et al., 2022). This raises a question of how mutant ATXN1 in astrocytes impacts their function, contributes to neuronal pathology, and leads to SCA1-like disease outcomes. To fill this knowledge gap, we aim to determine the molecular and cellular mechanisms though which mATXN1 affects cerebellar astrocytes, and how astrocyte dysfunctions contribute to the behavioral and pathological changes that are observed in SCA1 disease onset and progression.

NIH Research Projects · FY 2024 · 2024-07

ABSTRACT Cervical spine abnormalities in mucopolysaccharidosis (MPS) may result in cervical spinal cord (CSC) compression and irreversible neurological disability. MRI signal intensity changes in CSC images occur in the presence of advanced stenosis, myelopathy, and developed clinical symptoms. The proposal presumes that deficits in CSC morphology and microstructure precede MRI signal changes and are predictive of clinical myelopathy. We will proceed with systematic validation comparing CSC morphology and microstructure in MPS patients and case-matched healthy controls. The overall objective is to analyze existing and acquired CSC MRI data and assess longitudinal CSC morphology and cross-sectional CSC microstructure in the MPS. The central hypothesis is that quantitative MRI is sensitive to CSC morphological and microstructural degeneration, myelopathy risk, and disease severity in MPS. To test the central hypothesis and, thereby, attain the overall objective, we will pursue two specific aims: Aim 1: Assess longitudinal morphological damage of CSC in MPS types I, II, and VI and Aim 2: Assess cross-sectional microstructural damage of CSC in MPS types I and VI. Aim 1 will test a working hypothesis that CSC morphology is altered due to cervical spinal compression and is predictive of myelopathy in MPS. Aim 2 will test a working hypothesis that CSC inner integrity is altered due to MPS I and VI diseases. We will demonstrate that quantitative CSC MRI measurements are proportional to myelopathy risk and disease severity in MPS. The effects of MPS treatment on CSC measurements will be investigated where possible. The proposal will identify candidates for biomarkers of myelopathy in MPS. Regarding the biomarker research, the proposal is a pilot preclinical exploratory phase 1. The project is innovative because it will: (i) discover the biomarker candidates of myelopathy risk and disease severity in MPS; (ii) diffusion tensor imaging (DTI) will be assessed in the CSC of MPS patients; (iii) demonstrate that DTI-based CSC microstructure can assess ongoing degenerative processes before visual and qualitative MRI signal intensity changes. Overall, the proposal aims to advance objective early myelopathy diagnosis, which can guide the timing of a prophylactic myelopathy treatment and improve clinical outcomes in MPS.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY The supramammillary area (SuM) is increasingly appreciated for its role in a variety of functions, including spatial and social novelty processing via its connections with the hippocampus. Specifically, the SuM has projections to the CA2 and ventral dentate gyrus (DG), believed to be important for social processing, and to the dorsal DG, believed to be important for spatial and contextual processing. The hippocampus provides direct inhibitory input to the SuM, but the nature and function of this connectivity is completely unknown. The studies outlined in the proposal would provide important basic science groundwork for understanding this connectivity, with ultimate potential impacts for better understanding and treating a range of neurological disorders, including but not limited to temporal lobe epilepsy. This includes examining the hippocampal neurons providing the inhibitory input to the SuM, examining which subpopulations of SuM neurons receive inhibitory hippocampal input, and the in vivo impact of manipulation of inhibitory hippocampal inputs to the SuM. Based on our preliminary data, we predict that there are at least two populations of hippocampal inhibitory neurons projecting to the SuM -- one expressing neuronal nitric oxide synthase (nNOS) and the other expressing somatostatin – located in each region and along the entire anterior-posterior axis of the hippocampus. We further predict the greatest number in the CA3 region. We predict, and our preliminary data supports, that both SuM cells projecting to the ventral DG and SuM cells projecting to the dorsal DG receive inhibitory hippocampal inputs. Importantly, we see the strongest connectivity so far from the ventral hippocampus to SuM neurons projecting to the dorsal DG, arguing against a simple feedback inhibitory role of this connection. To examine the function of inhibitory hippocampal inputs to the SuM in vivo, we will use fiberphotometry to record from projection-defined SuM populations, during an assortment of behavioral tests, while providing closed-loop optogenetic manipulation of inhibitory hippocampal inputs specifically at the time of object or conspecific investigations. Collectively, the data will provide important insights into the nature of hippocampal inhibitory inputs to the SuM, including the degree to which specific SuM populations are targeted, and the functional significance of this connectivity, including as relates to social and spatial processing.

NIH Research Projects · FY 2024 · 2024-07

Project Summary The purpose of this R13 conference grant is to seek partial funding support for the second international Computational Psychiatry Conference. This meeting will be held over three days (16-18 July 2024) in Minneapolis, Minnesota on the campus of the University of Minnesota, providing a centralized location that is both accessible and compact, allowing for interactions among attendees. The conference builds on a very successful first iteration held in Dublin, Ireland in 2023, as well as a 6 year (2014-2019, 2022) successful workshop run out of University College London and then Mount Sinai medical school. The specific goal of the conference is to bring together early-career and established investigators working on clinical and computational questions related to mental health and addiction. Our aim is to build a productive and career-enhancing annual conference that supports the development of researchers to translate neuroscience discoveries into psychiatric clinical practice and to use clinical observations to generate new discoveries in neuroscience. The specific aim of this R13 proposal is to provide support for early career investigators for the following activities: ● Career development sessions, including ○ Curated interactions with to more senior investigators; ○ A panel discussion on diversity, equity, and inclusion in the computational psychiatry field; ○ A career journey discussion. ● Two tutorials to allow researchers with limited expertise to broaden their knowledge base; the tutorials are particularly important for this community because this conference brings together researchers from two very separate expertises (computational neuroscience and clinical psychiatry). ● Poster awards for trainees. ● Travel support for early career investigators from communities underrepresented in medicine and science.